Figure 3

Flow cytometric and functional analysis of major cellular subsets amplified in HSC expansion conditions with UM171. (A) Representative dot plots show phenotype of major subsets of CD34⁺CD43⁺ cells identified in cultures after 3 days of expansion and gates used for their sorting (CD235a vs CD41a dot plots on the left). (B) Bar depicts mean  ± SEM percentage of each subset within the CD34⁺CD43⁺ population at day 3 (n = 4) and day 5 (n = 7) of expansion. (C) CFC composition for each sorted population. Results are mean ± SEM for 6 independent experiments. (D) Identification and characterization of CD41a^lo population within CD45⁺ hematopoietic progenitors formed on day 9 of differentiation, before initiation of expansion. Dot plots show gates used for analysis. Pie graphs show relative proportions of each type of CFC within sorted populations. (E) Representative dot plots show phenotype of BrdU⁺ proliferating cells, after gating CD45⁺ cells from DMSO and UM171 expanded HPs. (F) Percentages of CD41a⁻, CD41a^lo, and CD41^hi subsets within BrdU⁺ proliferating cells after gating on the CD34⁺CD45⁺ population. Results are mean ± SEM (n = 3). **p < 0.01, ***p < 0.001. (G) Percentages of BrdU⁺ and BrdU⁻ cells within indicated subsets after 5 days of expansion; Results are mean ±  SEM (n = 3). *p < 0.05.